Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
DNA Topoisomerases02:02

DNA Topoisomerases

Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
Types and Mechanism of action
Topoisomerases are divided into two main types.  Type I...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

DMPK 3' untranslated repeat expansions in unexplained sudden cardiac death in the young.

European heart journal·2026
Same author

Effect of Glucoraphanin on the Abundance of Nrf2 Regulated Genes Within Circulating Small Extracellular Vesicles: A Pilot Dietary Intervention.

Molecular nutrition & food research·2026
Same author

Early termination of NCT04617067, a phase II, open label, clinical trial of oral paricalcitol in combination with gemcitabine and NAB-paclitaxel therapy in advanced pancreatic cancer.

BMC cancer·2026
Same author

Complete genome sequence of <i>Escherichia marmotae</i> F12YCO47 isolated from a healthy human fecal sample.

Microbiology resource announcements·2025
Same author

Benchmarking nanopore sequencing and rapid genomics feasibility: validation at a quaternary hospital in New Zealand.

NPJ genomic medicine·2024
Same author

GPS-derived environmental exposures during pregnancy and early postpartum - Evidence from the madres cohort.

The Science of the total environment·2024

Related Experiment Video

Updated: Jun 22, 2026

Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death
13:54

Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death

Published on: October 21, 2012

Studying vertebrate topoisomerase 2 function using a conditional knockdown system in DT40 cells.

Mark Johnson1, Hui Hui Phua, Sophia C Bennett

  • 1Department of Genetics, University of Cambridge, Downing St, Cambridge CB2 3EH, UK.

Nucleic Acids Research
|June 5, 2009
PubMed
Summary

Researchers developed a new method using RNA interference in DT40 cells to study topoisomerase 2 (topo 2) function. This technique allows for conditional depletion of topo 2 isoforms, enabling deeper insights into cell cycle regulation and DNA decatenation checkpoints.

More Related Videos

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique
07:18

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Published on: October 27, 2011

In Vivo Functional Study of Disease-associated Rare Human Variants Using Drosophila
06:41

In Vivo Functional Study of Disease-associated Rare Human Variants Using Drosophila

Published on: August 20, 2019

Related Experiment Videos

Last Updated: Jun 22, 2026

Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death
13:54

Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death

Published on: October 21, 2012

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique
07:18

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Published on: October 27, 2011

In Vivo Functional Study of Disease-associated Rare Human Variants Using Drosophila
06:41

In Vivo Functional Study of Disease-associated Rare Human Variants Using Drosophila

Published on: August 20, 2019

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • The DT40 avian B-cell line is valuable for studying gene function due to high homologous recombination rates.
  • Gene targeting is a common method, but RNA interference offers an alternative for gene function studies.

Purpose of the Study:

  • To develop and characterize a DT40 cell line for inducible knockdown of topoisomerase 2 (topo 2) isoforms using RNA interference.
  • To investigate the cell cycle effects of topo 2 depletion in DT40 cells.
  • To explore the role of topo 2 depletion in activating the G2 catenation checkpoint.

Main Methods:

  • In situ tagging of both topo 2 genes in DT40 cells using gene targeting.
  • Generation of stable transfectants with tetracycline-inducible short hairpin RNAs for conditional mRNA depletion of topo 2 isoforms.
  • Comparison of cell cycle phenotypes with previously published data from other cell types.

Main Results:

  • Successful generation of DT40 cells with tagged topo 2 genes and inducible knockdown capabilities.
  • Characterization of the cell cycle phenotype of topo 2-depleted DT40 cells.
  • Data suggests the DT40 knockdown system can be used to probe the G2 catenation checkpoint.

Conclusions:

  • RNA interference in DT40 cells provides a powerful alternative to gene targeting for studying essential gene functions.
  • The developed system allows for conditional depletion of topo 2 isoforms, facilitating detailed cell cycle analysis.
  • This approach is suitable for investigating DNA decatenation checkpoints and their role in cell cycle regulation.